Study on in vitro Cell Culture and Transgene for Sheep Fetal Fibroblasts
  Revised:May 26, 2003
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KeyWord:Enhanced green fluorescent protein,Sheep fetal fibroblasts,Single cell cloning,Transgenes
MA Yu-zhen,HU Ting-mao,WANG Jian-guo,HU Hui-ping,LIU Ming-qiu
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      Objective Sheep fetal fibroblasts(SFF) were transfected by enhanced green fluorescent protein(EGFP) gene to investigate the effect of the foreign gene on the biological characteristics of SFF. Methods SFF were dispersed and cultured in vitro.pEGFP-C1 was transfected to the passage 1 SFF by liposomes.The SFF were incubated in medium containing G418 for 10-12 days.Resistant colonies were picked and subcultured until use.The SFF were examined by cell morphology,growth curve or the nuclear type of chromosome.Sex-determining region Y gene(SRY) of the SFF was identified.Results The biological characteristics did not differ significantly between the resistant colonies and the control cells.Expressive amount of foreign gene was revealed by fluorescent intensity.Conclusion The present research can provide a support for the transgenic cell with EGFP as the report gene in vitro.Transgenic donor cells integrated with EGFP gene provide an efficient new strategy for the production of transgenic farm animals.
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